[go: up one dir, main page]

CN104706626B - A kind of microneedle patch for being easy to animal vaccine to be administered and preparation method thereof - Google Patents

A kind of microneedle patch for being easy to animal vaccine to be administered and preparation method thereof Download PDF

Info

Publication number
CN104706626B
CN104706626B CN201510131662.7A CN201510131662A CN104706626B CN 104706626 B CN104706626 B CN 104706626B CN 201510131662 A CN201510131662 A CN 201510131662A CN 104706626 B CN104706626 B CN 104706626B
Authority
CN
China
Prior art keywords
vaccine
microneedle
microneedle patch
patch
preparation
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN201510131662.7A
Other languages
Chinese (zh)
Other versions
CN104706626A (en
Inventor
郭新东
王淇磊
陈洋
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Beijing University of Chemical Technology
Original Assignee
Beijing University of Chemical Technology
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Beijing University of Chemical Technology filed Critical Beijing University of Chemical Technology
Priority to CN201510131662.7A priority Critical patent/CN104706626B/en
Publication of CN104706626A publication Critical patent/CN104706626A/en
Application granted granted Critical
Publication of CN104706626B publication Critical patent/CN104706626B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61MDEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M37/00Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin
    • A61M37/0015Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin by using microneedles
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61MDEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M37/00Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin
    • A61M37/0015Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin by using microneedles
    • A61M2037/0046Solid microneedles
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61MDEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M37/00Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin
    • A61M37/0015Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin by using microneedles
    • A61M2037/0053Methods for producing microneedles

Landscapes

  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Dermatology (AREA)
  • Medical Informatics (AREA)
  • Anesthesiology (AREA)
  • Biomedical Technology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Hematology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Medicinal Preparation (AREA)

Abstract

本发明属于给药设备技术领域,具体来说,涉及一种便于动物疫苗给药的微针贴片及其制备方法。所述微针贴片包括可溶性的贴片基底和阵列分布在贴片基底上的可溶性的微针针体;所述微针针体在其顶端载有目标药物,该微针针体的制备材料包括由水溶性的生物可降解的高分子材料。与现有技术相比,本发明所述的便于动物疫苗给药的微针贴片在接种疫苗时不会引起动物的惧怕反应,易于给药,操作方便,不需要对接种人员培训;单人即可完成接种疫苗工作,效率高;完成疫苗接种时不需额外对动物进行标记;一次性使用,避免了交叉感染的可能性;所用材料可生物降解,无医疗垃圾产生;制作方法简单经济;产品无需冷藏保存或真空保存等;运输方便。

The invention belongs to the technical field of drug delivery equipment, and in particular relates to a microneedle patch which is convenient for animal vaccine drug delivery and a preparation method thereof. The microneedle patch includes a dissolvable patch base and dissolvable microneedle bodies distributed in an array on the patch base; Including water-soluble biodegradable polymer materials. Compared with the prior art, the microneedle patch of the present invention that facilitates the administration of animal vaccines does not cause fear reactions in animals when vaccinated, is easy to administer, is convenient to operate, and does not require training for vaccinators; The vaccination can be completed with high efficiency; no additional marking of animals is required when the vaccination is completed; one-time use avoids the possibility of cross-infection; the materials used are biodegradable and no medical waste is generated; the production method is simple and economical; The product does not need to be refrigerated or vacuum stored, etc.; it is convenient to transport.

Description

一种便于动物疫苗给药的微针贴片及其制备方法A kind of microneedle patch convenient for animal vaccine administration and preparation method thereof

技术领域technical field

本发明属于给药设备技术领域,具体来说,涉及一种便于动物疫苗给药的微针贴片及其制备方法。The invention belongs to the technical field of drug delivery equipment, and in particular relates to a microneedle patch which is convenient for animal vaccine drug delivery and a preparation method thereof.

背景技术Background technique

疫苗是用以控制某一传染性疾病的重要武器。动物疫苗属于兽用生物制品的范畴,兽用生物制品是根据免疫学原理,利用微生物、寄生虫及其代谢产物或免疫应答产物制备的一类物质。这类物质专供动物相应疾病的诊断、治疗或预防之用。动物疫苗的发明与推广很好的预防、控制动物传染病的发生与流行,为我国畜牧业的健康发展作出了巨大贡献。Vaccines are an important weapon to control an infectious disease. Animal vaccines belong to the category of veterinary biological products. Veterinary biological products are a class of substances prepared from microorganisms, parasites and their metabolites or immune response products based on the principles of immunology. Such substances are intended exclusively for the diagnosis, treatment or prevention of corresponding diseases in animals. The invention and promotion of animal vaccines can prevent and control the occurrence and prevalence of animal infectious diseases, and have made great contributions to the healthy development of animal husbandry in my country.

动物疫苗的传统给药方法是胃肠外注射。使用注射器的肌肉注射是将药物通过刺入皮肤的注射针把药物输送到肌肉组织,扩散后被肌体吸收,引起免疫反应。传统注射方式注射疫苗时,会存在很多问题:由于注射引起的疼痛导致动物的惧怕反应,造成注射不便;注射时造成皮肤和肌肉的伤害或造成皮肤感染;注射时有断针的危险;有交叉感染的可能性;为避免交叉感染针头需要更换,一次性注射器带来的生物垃圾造成环境污染和资源浪费;规范的操作需要对操作人员进行培训;注射时一般需要多人配合,效率低下;注射完成后为避免混淆要对注射完成的动物进行标记等等。The traditional method of administration of animal vaccines is parenteral injection. Intramuscular injection using a syringe is to deliver the drug to the muscle tissue through the injection needle piercing the skin, and then diffuse and absorb it into the body, causing an immune response. When injecting vaccines by traditional injection methods, there will be many problems: the pain caused by the injection will cause the animal's fear reaction, which will cause inconvenient injection; the skin and muscle damage or skin infection will be caused during the injection; the needle will break during the injection; there will be crossover. The possibility of infection; in order to avoid cross-infection needles need to be replaced, and the biological waste brought by disposable syringes causes environmental pollution and waste of resources; standardized operations require training for operators; injections generally require the cooperation of multiple people, which is inefficient; injections After completion, to avoid confusion, the animals that have been injected should be marked, etc.

目前可替代的疫苗接种方法主要有透皮给药技术。透皮给药是在皮肤表面给药,使药物以接近恒定速度通过皮肤各层,经毛细血管吸收进入体循环产生全身或局部治疗作用。透皮疫苗给药属于非侵害性疫苗给药,依赖于药物通过皮肤的扩散。由于皮肤表面角质层的保护和屏障作用,药物经皮扩散的效率很低,用于疫苗有给药的有效性数据不够充分。At present, alternative vaccination methods mainly include transdermal delivery technology. Transdermal administration is administration on the surface of the skin, so that the drug passes through each layer of the skin at a nearly constant speed, and is absorbed into the systemic circulation through capillaries to produce systemic or local therapeutic effects. Transdermal vaccine delivery is a non-invasive vaccine delivery that relies on the diffusion of the drug through the skin. Due to the protection and barrier function of the stratum corneum on the skin surface, the efficiency of transdermal drug diffusion is very low, and the effectiveness data for vaccine administration is insufficient.

发明内容Contents of the invention

为解决上述技术问题,本发明提供了一种便于动物疫苗给药的微针贴片及其制备方法。In order to solve the above technical problems, the present invention provides a microneedle patch which is convenient for administering animal vaccines and a preparation method thereof.

本发明所述的一种便于动物疫苗给药的微针贴片,所述微针贴片包括可溶性的贴片基底和阵列分布在贴片基底上的可溶性的微针针体;所述微针针体在其顶 端载有目标药物,该微针针体的制备材料包括由水溶性的生物可降解的高分子材料。A microneedle patch that is convenient for animal vaccine administration according to the present invention, the microneedle patch includes a soluble patch base and a dissolvable microneedle needle body array distributed on the patch base; the microneedle The tip of the needle body is loaded with target medicine, and the preparation material of the microneedle body includes water-soluble biodegradable macromolecule material.

本发明所述的一种便于动物疫苗给药的微针贴片,所述高分子材料为聚乙烯醇、聚羧甲基纤维素、硫酸软骨素、聚乳酸聚乙醇酸共聚物、蚕丝蛋白、糊精、透明质酸、明胶中一种或几种。A microneedle patch that is convenient for animal vaccine administration according to the present invention, the polymer material is polyvinyl alcohol, polycarboxymethyl cellulose, chondroitin sulfate, polylactic acid polyglycolic acid copolymer, silk protein, One or more of dextrin, hyaluronic acid, and gelatin.

本发明所述的一种便于动物疫苗给药的微针贴片,所述动物疫苗为蓝耳疫苗、猪瘟疫苗、口蹄疫疫苗、犬咳疫苗、犬六联疫苗、狂犬疫苗、牛流行热疫苗、炭疽疫苗、牛布鲁氏杆菌疫苗,羊痘苗、伪狂犬病疫苗、猪瘟兔化弱毒疫苗、猪肺疫弱毒疫苗、猪丹毒氢氧化铝菌苗、仔猪红痢菌苗、猪链球菌氢氧化铝菌苗、猪喘气病疫苗中的一种或几种。A microneedle patch that is convenient for administration of animal vaccines according to the present invention, the animal vaccines are blue ear vaccine, swine fever vaccine, foot-and-mouth disease vaccine, canine cough vaccine, canine six-in-one vaccine, rabies vaccine, bovine epidemic fever vaccine , anthrax vaccine, bovine brucellosis vaccine, sheep pox vaccine, pseudorabies vaccine, attenuated swine fever vaccine, porcine pneumonia attenuated vaccine, swine erysipelas aluminum hydroxide vaccine, piglet red dysentery vaccine, streptococcus suis hydrogenated One or more of aluminum bacterin vaccine and porcine asthma vaccine.

本发明所述的一种便于动物疫苗给药的微针贴片,所述制备材料还包括稳定剂,该稳定剂为蔗糖、葡萄糖、红菇酮萜、海藻糖、壳聚糖中一种或几种。A microneedle patch that is convenient for animal vaccine administration according to the present invention, the preparation material also includes a stabilizer, the stabilizer is one or Several kinds.

本发明所述的一种便于动物疫苗给药的微针贴片,所述目标药物的载药方式为将药物涂抹在微针针体顶端的外表、混匀在微针针体顶端或集中在微针针体顶端。A microneedle patch that is convenient for animal vaccine administration according to the present invention, the drug loading method of the target drug is to smear the drug on the surface of the top of the microneedle body, mix it on the top of the microneedle body or concentrate it on the surface of the microneedle body The tip of the microneedle body.

本发明所述的一种便于动物疫苗给药的微针贴片,所述微针贴片是边长为2-50mm的方形,共有1-10000个微针,相邻微针之间的间距是0.2-5mm。A microneedle patch that is convenient for administering animal vaccines according to the present invention, the microneedle patch is a square with a side length of 2-50 mm, with 1-10000 microneedles in total, and the distance between adjacent microneedles It is 0.2-5mm.

本发明所述的一种便于动物疫苗给药的微针贴片,所述微针针体的形状为圆锥形或类圆锥形,该微针针体长度为200-3000μm,底径为100-1500μm。A microneedle patch that is convenient for animal vaccine administration according to the present invention, the shape of the microneedle body is conical or quasi-conical, the length of the microneedle body is 200-3000 μm, and the bottom diameter is 100- 1500μm.

本发明所述的一种便于动物疫苗给药的微针贴片的制备方法,所述制备方法包括以下步骤:按质量百分比将5%-50%的高分子材料、0%-50%的稳定剂与余量的水混合,在40℃-100℃下搅拌溶解;溶解完全后抽真空去除气泡,得到高分子材料/糖的高浓度溶液,密封;按计量要求,配制浓度范围0.1-1000mg/ml的疫苗溶液;取疫苗溶液0.01-5ml涂在微针模板上,抽真空0.5-60min后,回收多余的疫苗溶液,继续在真空环境干燥0.5-60min,待疫苗溶液沉积到微针顶端;取高浓度溶液0.01-5ml涂到上述载有疫苗溶液的微针模板上,再次抽真空1-300min;冷冻干燥5-600min,干燥后脱模,在温度为15-25℃,湿度10%-30%RH下的密闭空间中进行封装,得微针贴片。A method for preparing a microneedle patch that is convenient for animal vaccine administration according to the present invention, the preparation method includes the following steps: 5%-50% of polymer materials, 0%-50% of stable Mix the agent with the remaining amount of water, stir and dissolve at 40°C-100°C; after the dissolution is complete, vacuumize to remove air bubbles to obtain a high-concentration solution of polymer material/sugar, and seal it; according to the measurement requirements, the concentration range of 0.1-1000mg/ ml of vaccine solution; take 0.01-5ml of vaccine solution and apply it on the microneedle template. Apply 0.01-5ml of the high-concentration solution on the above-mentioned microneedle template loaded with the vaccine solution, vacuumize again for 1-300min; Encapsulate in a closed space under %RH to obtain a microneedle patch.

与现有技术相比,本发明所述的便于动物疫苗给药的微针贴片在接种疫苗时 不会引起动物的惧怕反应,易于给药,操作方便,不需要对接种人员培训;单人即可完成接种疫苗工作,效率高;完成疫苗接种时不需额外对动物进行标记;一次性使用,避免了交叉感染的可能性;所用材料可生物降解,无医疗垃圾产生;制作方法简单经济;产品无需冷藏保存或真空保存等;运输方便。Compared with the prior art, the microneedle patch of the present invention that facilitates the administration of animal vaccines does not cause fear reactions in animals when vaccinated, is easy to administer, is convenient to operate, and does not require training for vaccinators; The vaccination can be completed with high efficiency; no additional marking of animals is required when the vaccination is completed; one-time use avoids the possibility of cross-infection; the materials used are biodegradable and no medical waste is generated; the production method is simple and economical; The product does not need to be refrigerated or vacuum stored, etc.; it is convenient to transport.

附图说明Description of drawings

图1:微针模板示意图;图2:微针贴片制备流程图;图3:微针贴片示意图;图4:微针贴片侧面示意图;图5:微针贴片显微图;图6:微针贴片扎猪皮实验显微图。Figure 1: Schematic diagram of microneedle template; Figure 2: Flow chart of preparation of microneedle patch; Figure 3: Schematic diagram of microneedle patch; Figure 4: Schematic diagram of the side of microneedle patch; Figure 5: Micrograph of microneedle patch; Fig. 6: Micrograph of pig skin piercing experiment with microneedle patch.

具体实施方式detailed description

下面结合具体的实施例对本发明所述的便于动物疫苗给药的微针贴片做进一步说明,但是本发明的保护范围并不限于此。The microneedle patch that facilitates the administration of animal vaccines according to the present invention will be further described below in conjunction with specific examples, but the scope of protection of the present invention is not limited thereto.

实施例1Example 1

一种便于动物疫苗给药的微针贴片,所述微针贴片包括可溶性微针基底和可溶性微针针体;所述可溶性微针针体载有目标药物,其制备材料包括由水溶性生物可降解的高分子材料;所述高分子材料为重均分子量为2000-300000的聚乙烯醇;所述制备材料还包括稳定剂,该稳定剂为蔗糖;所述目标药物的载药方式为尖端载药;所述微针贴片是边长为10mm的方形,共有100个微针,相邻微针之间的间距是0.5mm;所述微针针体的形状为圆锥形或类圆锥形,该微针针体长度为700μm,底径为500μm。A microneedle patch that facilitates the administration of animal vaccines, the microneedle patch includes a soluble microneedle base and a soluble microneedle body; the soluble microneedle body is loaded with a target drug, and its preparation material includes a water-soluble A biodegradable polymer material; the polymer material is polyvinyl alcohol with a weight average molecular weight of 2000-300000; the preparation material also includes a stabilizer, which is sucrose; the drug loading method of the target drug is The tip is loaded with medicine; the microneedle patch is a square with a side length of 10 mm, and there are 100 microneedles in total, and the distance between adjacent microneedles is 0.5 mm; the shape of the microneedle body is conical or cone-like shape, the length of the microneedle body is 700 μm, and the bottom diameter is 500 μm.

本发明所述的一种便于动物疫苗给药的微针贴片的制备方法,所述制备方法包括以下步骤:取50mg磺酰罗丹明(紫红色,在荧光照射下显亮红色,以此作为可溶性微针的模型药物)配制1mg/ml的磺酰罗丹明溶液;取18g聚乙烯醇、12g蔗糖、30g水混合,在90℃下加热搅拌,完全溶解后抽真空去除气泡,制得聚乙烯醇质量分数为30%,蔗糖质量分数为20%的高浓度溶液;将磺酰罗丹明溶液涂在微针模板上,抽真空15min后,回收多余的磺酰罗丹明溶液,继续在真空环境干燥20min,待磺酰罗丹明溶液沉积到微针顶端;将高浓度溶液覆盖到上述载有磺酰罗丹明溶液的微针模板上(注:贴片基底会与微针针体的下段一块成形),回收多余的高浓度溶液,再次抽真空1h后,放置于20℃通风橱干燥上,固化成型后将其转移到厚度1mm的贴片上,再次冷冻干燥5h脱水,在温度为20℃,湿度20%RH 下的密闭空间中进行封装,得微针贴片。A kind of preparation method of the microneedle patch that is convenient to animal vaccine administration according to the present invention, described preparation method comprises the following steps: take 50mg sulforhodamine (purple, bright red under fluorescent irradiation, use this as Soluble microneedle model drug) to prepare 1mg/ml sulforhodamine solution; mix 18g polyvinyl alcohol, 12g sucrose, and 30g water, heat and stir at 90°C, vacuumize to remove air bubbles after complete dissolution, and obtain polyethylene The mass fraction of alcohol is 30%, the mass fraction of sucrose is a high concentration solution of 20%; the sulforhodamine solution is coated on the microneedle template, and after vacuuming for 15 minutes, the excess sulforhodamine solution is recovered and dried in a vacuum environment After 20 minutes, the sulforhodamine solution is deposited on the top of the microneedle; cover the high-concentration solution on the above-mentioned microneedle template loaded with the sulforhodamine solution (note: the base of the patch will be formed together with the lower section of the microneedle body) , recover the excess high-concentration solution, vacuum again for 1 hour, place it on a fume hood at 20°C for drying, and transfer it to a patch with a thickness of 1mm after solidification and molding, and then freeze-dry for 5 hours to dehydrate. Packaged in a closed space under 20% RH to obtain a microneedle patch.

本实施例中得到的微针贴片在光学显微镜下可以清晰地观察到100个针体,单个贴片中的每个微针都具有锐利尖部,呈类圆锥状,尺寸均一;每个微针针体前端都有紫红色药物堆积,顶端颜色最深,而微针尖端以下部分和基座部分并没有紫红色药物。在荧光显微镜下可清晰地观察到微针尖端部分有亮红色荧光,针体其他部分和微针基座部分无红色荧光。以上光学显微镜和荧光显微镜观察到的现象表明,模型磺酰罗丹明溶液很好地载负到了微针针体尖端部分,针体其他部分和微针基座部分并无药物。The microneedle patch obtained in this example can clearly observe 100 needles under an optical microscope, and each microneedle in a single patch has a sharp tip, which is conical in shape and uniform in size; each microneedle There are purplish red drug accumulations at the tip of the needle body, and the top is the darkest, but there is no purplish red drug in the part below the tip of the microneedle and the base. Under the fluorescence microscope, it can be clearly observed that there is bright red fluorescence at the tip of the microneedle, but there is no red fluorescence at the other parts of the needle body and the base of the microneedle. The above phenomena observed by the optical microscope and the fluorescence microscope show that the model sulforhodamine solution is well loaded to the tip of the microneedle, and there is no drug in other parts of the needle and the base of the microneedle.

实施例2Example 2

一种便于动物疫苗给药的微针贴片,所述微针贴片包括可溶性微针基底和可溶性微针针体;所述可溶性微针针体载有目标药物,其制备材料包括由水溶性生物可降解的高分子材料;所述高分子材料为重均分子量为2000-300000的聚乙烯醇;所述制备材料还包括稳定剂,该稳定剂为红菇酮萜;所述目标药物的载药方式为尖端载药;所述微针贴片是边长为10mm的方形,共有100个微针,相邻微针之间的间距是0.7mm;所述微针针体的形状为圆锥形或类圆锥形,该微针针体长度为700μm,底径为500μm。A microneedle patch that facilitates the administration of animal vaccines, the microneedle patch includes a soluble microneedle base and a soluble microneedle body; the soluble microneedle body is loaded with a target drug, and its preparation material includes a water-soluble A biodegradable polymer material; the polymer material is polyvinyl alcohol with a weight-average molecular weight of 2000-300000; the preparation material also includes a stabilizer, which is rubrumone terpene; the loading of the target drug The way of medicine is drug loading on the tip; the microneedle patch is a square with a side length of 10mm, and there are 100 microneedles in total, and the distance between adjacent microneedles is 0.7mm; the shape of the microneedle body is conical Or conical shape, the length of the microneedle body is 700 μm, and the bottom diameter is 500 μm.

本发明所述的一种便于动物疫苗给药的微针贴片的制备方法,所述制备方法包括以下步骤:取猪蓝耳病疫苗与疫苗佐剂配制疫苗质量百分数为0.5%的疫苗溶液;取18g聚乙烯醇、12g红菇酮萜、30g水混合,在90℃下加热搅拌,完全溶解后抽真空去除气泡,制得聚乙烯醇质量分数为30%,红菇酮萜质量分数为20%的高浓度溶液;将疫苗溶液涂在微针模板上,抽真空60min后,回收多余的疫苗溶液,继续在真空环境干燥60min,待疫苗溶液沉积到微针顶端;将高浓度溶液覆盖到上述载有疫苗溶液的微针模板上(注:贴片基底会与微针针体的下段一块成形),回收多余的高浓度溶液,再次抽真空5h后,放置于20℃通风橱干燥上,固化成型后将其转移到厚度1mm的贴片上,再次冷冻干燥5h脱水,在温度为25℃,湿度30%RH下的密闭空间中进行封装,得微针贴片。经过比较,相对于蔗糖、葡萄糖、海藻糖及壳聚糖,采用从臭红菇中分离出的红菇酮萜为稳定剂,所制得的微针贴片其疫苗活性至少要高出12.4%,但其成本至少高出4倍,目前还不宜常规化。A method for preparing a microneedle patch that is convenient for animal vaccine administration according to the present invention, the preparation method includes the following steps: taking the porcine PRRS vaccine and vaccine adjuvant to prepare a vaccine solution with a mass percentage of 0.5% of the vaccine; Take 18g of polyvinyl alcohol, 12g of russula ketone, and 30g of water, mix them, heat and stir at 90°C, and vacuumize to remove air bubbles after completely dissolving, so that the mass fraction of polyvinyl alcohol is 30%, and the mass fraction of russula ketone is 20%. % high-concentration solution; apply the vaccine solution on the microneedle template, after vacuuming for 60 minutes, recover the excess vaccine solution, continue to dry in a vacuum environment for 60 minutes, and wait for the vaccine solution to deposit on the top of the microneedle; cover the high-concentration solution on the above-mentioned On the microneedle template loaded with the vaccine solution (note: the base of the patch will be formed together with the lower part of the microneedle body), recover the excess high-concentration solution, and after vacuuming again for 5 hours, place it on a fume hood at 20°C to dry and solidify After molding, transfer it to a patch with a thickness of 1 mm, freeze-dry it again for 5 hours to dehydrate, and package it in a closed space at a temperature of 25° C. and a humidity of 30% RH to obtain a microneedle patch. After comparison, compared with sucrose, glucose, trehalose and chitosan, the vaccine activity of the prepared microneedle patch is at least 12.4% higher than that of sucrose, glucose, trehalose and chitosan, using Russula ketone isolated from Stinky Russula as a stabilizer , but its cost is at least 4 times higher, and it is not suitable for routine use at present.

以10只体重约20kg的年龄在7周左右的雌性猪为实验对象,其中5只作为实 验对象接种疫苗,另外5只不接种作为对照组。将本实施例中得到的蓝耳病疫苗微针贴片按压在猪的左臀部,按压时尽量使微针垂直刺入猪皮肤,按压时间2分钟,使可溶性疫苗微针顶端的疫苗完全释放到猪皮肤中,完成经皮给药。两周后重复一次上述给药。并在第二次给药前一天从猪尾静脉抽血并获得血清,通过ELISA方法测定血清中针对蓝耳病疫苗的特异性IgG1抗体的滴度。实验结果是,对照组通过上述制备的蓝耳病疫苗微针贴片接种的实验猪血清中的特异性IgG1抗体的平均滴度滴度为6log2,对照组特异性IgG1抗体的平均滴度滴度为0log2。说明通过本发明所述的可溶性高分子微针贴片用于猪蓝耳病疫苗的给药成功,效果显著。Take 10 female pigs with a body weight of about 20kg and an age of about 7 weeks as the experimental subjects, 5 of which were vaccinated as the experimental subjects, and the other 5 were not vaccinated as the control group. Press the microneedle patch of the PRRS vaccine obtained in this example on the left buttocks of the pig, try to make the microneedle vertically pierce the pig's skin when pressing, and press for 2 minutes, so that the vaccine at the top of the soluble vaccine microneedle is completely released into the pig's skin. In porcine skin, transdermal administration was accomplished. The above administration was repeated two weeks later. And one day before the second administration, blood was drawn from pig tail vein to obtain serum, and the titer of specific IgG1 antibody against PRRS vaccine in serum was determined by ELISA method. The result of the experiment is that the average titer titer of the specific IgG1 antibody in the experimental pig serum inoculated by the PRRS vaccine microneedle patch prepared above is 6log2 for the control group, and the average titer titer of the specific IgG1 antibody for the control group is 6log2. is 0log2. It shows that the soluble polymer microneedle patch of the present invention is used for administration of porcine PRRS vaccine successfully, and the effect is remarkable.

实施例3Example 3

一种便于动物疫苗给药的微针贴片,所述微针贴片包括可溶性微针基底和可溶性微针针体;所述可溶性微针针体载有目标药物,其制备材料包括由水溶性生物可降解的高分子材料;所述高分子材料为重均分子量为2000-300000的聚乙烯醇;所述制备材料还包括稳定剂,该稳定剂为海藻糖;所述目标药物的载药方式为尖端载药;所述微针贴片是边长为10mm的方形,共有100个微针,相邻微针之间的间距是0.6mm;所述微针针体的形状为圆锥形或类圆锥形,该微针针体长度为600μm,底径为500μm。A microneedle patch that facilitates the administration of animal vaccines, the microneedle patch includes a soluble microneedle base and a soluble microneedle body; the soluble microneedle body is loaded with a target drug, and its preparation material includes a water-soluble A biodegradable polymer material; the polymer material is polyvinyl alcohol with a weight average molecular weight of 2000-300000; the preparation material also includes a stabilizer, which is trehalose; the drug loading method of the target drug The tip is loaded with medicine; the microneedle patch is a square with a side length of 10 mm, and there are 100 microneedles in total, and the distance between adjacent microneedles is 0.6 mm; the shape of the microneedle body is conical or similar Conical shape, the length of the microneedle body is 600 μm, and the bottom diameter is 500 μm.

本发明所述的一种便于动物疫苗给药的微针贴片的制备方法,所述制备方法包括以下步骤:取狂犬疫苗与疫苗佐剂配制疫苗质量百分数为0.5%的疫苗溶液;取18g聚乙烯醇、12g海藻糖、30g水混合,在90℃下加热搅拌,完全溶解后抽真空去除气泡,制得聚乙烯醇质量分数为30%,海藻糖质量分数为20%的高浓度溶液;将疫苗溶液涂在微针模板上,抽真空15min后,回收多余的疫苗溶液,继续在真空环境干燥20min,待疫苗溶液沉积到微针顶端;将高浓度溶液覆盖到上述载有疫苗溶液的微针模板上(注:贴片基底会与微针针体的下段一块成形),回收多余的高浓度溶液,再次抽真空1h后,放置于20℃通风橱干燥上,固化成型后将其转移到厚度1mm的贴片上,再次冷冻干燥0.5h脱水,在温度为15℃,湿度10%RH下的密闭空间中进行封装,得微针贴片。A preparation method of a microneedle patch that is convenient for animal vaccine administration according to the present invention, the preparation method comprises the following steps: taking rabies vaccine and vaccine adjuvant to prepare a vaccine solution with a vaccine mass percentage of 0.5%; taking 18g poly Vinyl alcohol, 12g trehalose, and 30g water were mixed, heated and stirred at 90°C, vacuumized to remove air bubbles after complete dissolution, and a high-concentration solution with a mass fraction of polyvinyl alcohol of 30% and a mass fraction of trehalose of 20% was obtained; Apply the vaccine solution on the microneedle template, after vacuuming for 15 minutes, recover the excess vaccine solution, continue to dry in a vacuum environment for 20 minutes, and wait for the vaccine solution to deposit on the top of the microneedle; cover the high-concentration solution on the microneedle loaded with the vaccine solution On the template (note: the base of the patch will be formed together with the lower part of the microneedle body), recover the excess high-concentration solution, vacuumize again for 1 hour, place it on a fume hood at 20°C to dry, and transfer it to the thickness of the microneedle after curing On the 1mm patch, freeze-dry again for 0.5h dehydration, and package in a closed space at a temperature of 15° C. and a humidity of 10% RH to obtain a microneedle patch.

以5只体重约10kg的年龄在7周左右的雌性犬为实验对象。将本实施例中得到的狂犬疫苗微针贴片按压在犬的左下腹部,按压时尽量使微针垂直刺入皮肤, 按压时间2分钟,使可溶性疫苗微针顶端的疫苗完全释放到犬皮肤中,完成经皮给药。两周后重复一次上述给药。并在第二次给药前一天从静脉抽血并获得血清,测定血清中针对链球菌的特异性IgG1抗体平均滴度。实验结果是,对照组通过上述制备的狂犬疫苗微针贴片接种的实验犬血清中的特异性IgG1抗体的平均滴度滴度为4log2,对照组特异性IgG1抗体的平均滴度滴度为0log2。说明通过本发明所述的可溶性高分子微针贴片用于狂犬疫苗给药成功,效果显著。Five female dogs weighing about 10 kg and aged about 7 weeks were used as the experimental subjects. Press the rabies vaccine microneedle patch obtained in this example on the left lower abdomen of the dog. When pressing, try to make the microneedle penetrate the skin vertically. The pressing time is 2 minutes, so that the vaccine at the top of the soluble vaccine microneedle is completely released into the dog's skin. , to complete transdermal administration. The above administration was repeated two weeks later. And one day before the second administration, blood was drawn from the vein and serum was obtained, and the average titer of specific IgG1 antibody against streptococcus in the serum was determined. The result of the experiment is that the average titer titer of the specific IgG1 antibody in the experimental dog serum vaccinated by the rabies vaccine microneedle patch prepared above for the control group is 4log2, and the average titer titer of the specific IgG1 antibody for the control group is 0log2 . It shows that the soluble polymer microneedle patch of the present invention is used for rabies vaccine administration successfully, and the effect is remarkable.

效果验证Effect verification

1、微针皮肤试验1. Microneedle skin test

预先准备未脱毛的平整的新鲜猪皮,取上述制得的载负荧光模型药物磺酰罗丹明溶液的聚乙烯醇微针贴片,将微针针尖对准猪皮表层垂直刺入,用手按压微针贴片的背面,按压的时间为2min,然后取下作用后的微针贴片,可在光学显微镜和荧光显微镜下观察作用后的猪皮。在光学显微镜下可观察到,作用后的猪皮表面有100个清晰的红色的针孔,其他部分皮肤表面完好,红色针孔为微针贴片的可溶性微针针体顶端载负的紫红色模型药物磺酰罗丹明溶液所留,表明微针贴片的可溶性微针针体可有效地刺穿猪皮表层,在皮下溶解并且将可溶性微针顶端载负的模型药物磺酰罗丹明溶液释放到猪皮之中。同样的,在荧光显微镜下可观察到,作用后的猪皮表面有100个清晰地发出红色荧光的针孔,其他部分显示黑色,说明红色荧光的针孔为微针贴片的可溶性微针针体顶端载负的紫红色模型药物磺酰罗丹明溶液所留,表明微针贴片的可溶性微针针体可有效地刺穿猪皮表层,在皮下溶解并且将可溶性微针顶端载负的模型药物磺酰罗丹明溶液释放到猪皮之中。Prepare undepilated flat fresh pigskin in advance, take the polyvinyl alcohol microneedle patch loaded with the fluorescent model drug sulforhodamine solution prepared above, align the tip of the microneedle with the surface of the pigskin and insert it vertically. Press the back of the microneedle patch for 2 minutes, then remove the microneedle patch, and observe the activated pigskin under an optical microscope and a fluorescent microscope. It can be observed under the optical microscope that there are 100 clear red pinholes on the surface of the pig skin after the treatment, and the other parts of the skin surface are intact. The red pinholes are the purple red on the top of the soluble microneedle body of the microneedle patch. The model drug sulforhodamine solution remains, indicating that the soluble microneedle body of the microneedle patch can effectively pierce the surface of pig skin, dissolve subcutaneously and release the model drug sulforhodamine solution loaded on the top of the soluble microneedle into the pigskin. Similarly, it can be observed under a fluorescent microscope that there are 100 pinholes clearly emitting red fluorescence on the surface of the treated pigskin, and the other parts are black, indicating that the red fluorescent pinholes are the soluble microneedle needles of the microneedle patch The purple-red model drug sulforhodamine solution loaded on the top of the body is left behind, indicating that the soluble microneedle body of the microneedle patch can effectively pierce the surface layer of pig skin, dissolve subcutaneously and load the top of the soluble microneedle A solution of the drug sulforhodamine was released into the pig skin.

2、微针皮下溶解动力学2. Microneedle subcutaneous dissolution kinetics

按照实施例1中的步骤制备200个微针贴片,每个微针贴片面积1cm2有10×10个微针,针体长度700μm,间距0.5mm;每个微针贴片载负模型药物磺酰罗丹明溶液的质量是0.2mg。每十个微针贴片一组,一共20组。按照实施例2中的皮肤试验步骤,将上述微针贴片插入相同实验条件的猪皮,每组微针按压时间不同,1组按压时间10s后取下,2组按压时间是20s,3组按压时间是30s,依次类推,第20组按压时间是200s。在皮肤实验之后,检测每一片给药之后的的猪皮肤上的有效给药率,即皮肤上沉积的药物占微针贴片总载药量的百分比,结果如表1所示。Prepare 200 microneedle patches according to the steps in Example 1, each microneedle patch has an area of 1 cm 2 with 10×10 microneedles, the length of the needle body is 700 μm, and the spacing is 0.5 mm; each microneedle patch is loaded with a model The mass of the drug sulforhodamine solution is 0.2 mg. Each set of ten microneedle patches, a total of 20 sets. According to the skin test procedure in Example 2, the above-mentioned microneedle patch was inserted into the pigskin under the same experimental conditions. The pressing time of the microneedles in each group was different. The pressing time of the first group was removed after 10 seconds, the pressing time of the second group was 20 seconds, and the pressing time of the third group was 20 seconds. The pressing time is 30s, and so on, the pressing time of the 20th group is 200s. After the skin test, the effective drug delivery rate on each piece of pig skin after administration was detected, that is, the percentage of the drug deposited on the skin to the total drug loading of the microneedle patch, and the results are shown in Table 1.

表1:不同按压时间的有效给药率Table 1: Effective drug delivery rate for different compression times

给药时间/sAdministration time/s 11 22 33 44 55 66 77 88 99 1010 有效给药率/%Effective drug delivery rate/% 30.530.5 42.142.1 48.348.3 59.359.3 65.765.7 68.968.9 72.772.7 7474 77.377.3 79.679.6 给药时间/sAdministration time/s 1111 1212 1313 1414 1515 1616 1717 1818 1919 2020 有效给药率/%Effective drug delivery rate/% 83.683.6 86.886.8 90.290.2 92.992.9 94.294.2 95.995.9 97.697.6 98.298.2 9999 99.6 99.6

与现有技术相比,本发明所述的便于动物疫苗给药的微针贴片在接种疫苗时不会引起动物的惧怕反应,易于给药,操作方便,不需要对接种人员培训;单人即可完成接种疫苗工作,效率高;完成疫苗接种时不需额外对动物进行标记;一次性使用,避免了交叉感染的可能性;所用材料可生物降解,无医疗垃圾产生;制作方法简单经济;产品无需冷藏保存或真空保存等;运输方便。Compared with the prior art, the microneedle patch of the present invention that facilitates the administration of animal vaccines does not cause fear reactions in animals when vaccinated, is easy to administer, is convenient to operate, and does not require training for vaccinators; The vaccination can be completed with high efficiency; no additional marking of animals is required when the vaccination is completed; one-time use avoids the possibility of cross-infection; the materials used are biodegradable and no medical waste is generated; the production method is simple and economical; The product does not need to be refrigerated or vacuum stored, etc.; it is convenient to transport.

Claims (3)

1. it is a kind of be easy to animal vaccine be administered microneedle patch preparation method, it is characterised in that the preparation method include with Lower step:5%-50% high polymer material, 0%-50% stabilizer are mixed with the water of surplus by mass percentage, 40 DEG C- Stirring and dissolving at 100 DEG C;Bubble removing is vacuumized after dissolving completely, obtains highly concentrated solution, is sealed;By measuring requirement, prepare Concentration range 0.1-1000mg/ml vaccine solution;Take vaccine solution 0.01-5ml to be coated on microneedles template, vacuumize 0.5- After 60min, unnecessary vaccine solution is reclaimed, continues to dry 0.5-60min in vacuum environment, treats that vaccine solution deposits to micropin top End;Take highly concentrated solution 0.01-5ml be coated onto it is above-mentioned be loaded with the microneedles template of vaccine solution, vacuumize 1-300min again;It is cold Dry 5-600min is freezed, is stripped after drying, is 15-25 DEG C in temperature, is sealed in the confined space under humidity 10%-30%RH Dress, obtains microneedle patch;
The microneedle patch includes the soluble micropin needle body of soluble paster substrate and array distribution in paster substrate; The micropin needle body is loaded with drug target on its top, and the micropin needle body prepares material including water miscible biodegradable High polymer material;The stabilizer is one or more of in sucrose, russule ketone terpene, trehalose;The high polymer material is poly- second It is one or more of in enol, chondroitin sulfate, hyaluronic acid;The animal vaccine is blue ear vaccine or rabies vaccine.
2. preparation method according to claim 1, it is characterised in that the microneedle patch is the side that the length of side is 2-50mm Shape, 1-10000 micropin is shared, the spacing between adjacent micropin is 0.2-5mm.
3. preparation method according to claim 1, it is characterised in that the micropin needle body is shaped as cone or class circle Taper, the micropin needle body length are 200-3000 μm, and bottom footpath is 100-1500 μm.
CN201510131662.7A 2015-03-25 2015-03-25 A kind of microneedle patch for being easy to animal vaccine to be administered and preparation method thereof Active CN104706626B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201510131662.7A CN104706626B (en) 2015-03-25 2015-03-25 A kind of microneedle patch for being easy to animal vaccine to be administered and preparation method thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201510131662.7A CN104706626B (en) 2015-03-25 2015-03-25 A kind of microneedle patch for being easy to animal vaccine to be administered and preparation method thereof

Publications (2)

Publication Number Publication Date
CN104706626A CN104706626A (en) 2015-06-17
CN104706626B true CN104706626B (en) 2018-02-23

Family

ID=53406675

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201510131662.7A Active CN104706626B (en) 2015-03-25 2015-03-25 A kind of microneedle patch for being easy to animal vaccine to be administered and preparation method thereof

Country Status (1)

Country Link
CN (1) CN104706626B (en)

Families Citing this family (19)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105078880A (en) * 2015-09-12 2015-11-25 北京化工大学 Macromolecular soluble microneedle used for cutaneous penetration of polypeptide and protein medicines and preparation method of macromolecular soluble microneedle
KR101610598B1 (en) 2015-09-21 2016-04-07 비엔엘바이오테크 주식회사 FLEXIBLE MICRONEEDLE FOR DENTAL MATERIAL DELIVERY AND THE MANUFACTURING METHOD Of THE SAME
US20180264244A1 (en) 2015-09-28 2018-09-20 Vaxxas Pty Limited Microprojection arrays with enhanced skin penetrating properties and methods thereof
CN105726458B (en) * 2016-01-29 2018-09-28 广州新济药业科技有限公司 Responsive to temperature type solubility micropin and preparation method thereof
CN105596287B (en) * 2016-02-04 2018-09-18 广州新济药业科技有限公司 Active divergence type solubility micropin and preparation method thereof
CN106039552A (en) * 2016-05-30 2016-10-26 北京化工大学 Bubble-type microneedle and preparation method therefor
CN106422045A (en) * 2016-09-05 2017-02-22 中国科学院理化技术研究所 Flexible slow-release microneedle patch and preparation method thereof
CN106474620A (en) * 2016-09-22 2017-03-08 北京化工大学 A kind of polymer micro needle of medicine controlled release, preparation method and microneedle patch
CN110709250B (en) 2017-03-31 2022-10-11 瓦克萨斯私人有限公司 Apparatus and method for coating a surface
TWI633901B (en) * 2017-05-16 2018-09-01 怡定興科技股份有限公司 Medical beauty microneedle patch manufacturing method
AU2018285954A1 (en) 2017-06-13 2019-12-19 Vaxxas Pty Limited Quality control of substrate coatings
CN108379573B (en) * 2018-03-26 2020-10-16 中国医学科学院生物医学工程研究所 Pertussis vaccine microneedle array and preparation method thereof
CN110680909B (en) * 2018-07-04 2024-09-20 辽宁成大生物股份有限公司 Quick-release type b haemophilus influenzae combined vaccine soluble microneedle patch and preparation method thereof
CN110840822B (en) * 2018-07-26 2021-05-18 华中科技大学 Method for preparing porous polymer microneedle and application thereof
CN110870943A (en) * 2018-08-31 2020-03-10 中科微针(北京)科技有限公司 Implantable two-section type microneedle patch and preparation method thereof
WO2020198785A1 (en) * 2019-03-29 2020-10-08 Vaxxas Pty Ltd Vaccination using high-density microprojection array patch
CN113368033A (en) * 2021-06-09 2021-09-10 中国人民解放军军事科学院军事医学研究院 Heat-stable soluble microneedle vaccine patch based on animal gelatin and preparation method thereof
CN114533649A (en) * 2022-01-05 2022-05-27 华中科技大学同济医学院附属协和医院 Separable microneedle array capable of drug delivery and in-situ labeling and preparation method thereof
CN120227361A (en) * 2023-12-29 2025-07-01 珠海科瑞微医药科技有限公司 Preparation process of microneedle patch and microneedle patch prepared by preparation process

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103568160A (en) * 2012-07-27 2014-02-12 中国科学院理化技术研究所 Method for manufacturing polymer material micro-needle array patch
CN104367567A (en) * 2014-10-22 2015-02-25 北京化工大学 Extrusion embossing based quick vaccine patch and preparation method thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103568160A (en) * 2012-07-27 2014-02-12 中国科学院理化技术研究所 Method for manufacturing polymer material micro-needle array patch
CN104367567A (en) * 2014-10-22 2015-02-25 北京化工大学 Extrusion embossing based quick vaccine patch and preparation method thereof

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
The development and characteristics of novel microneedle arrays fabricated from hyaluronic acid, and their application in the transdermal delivery of insulin;Shu Liu 等;《Journal of Controlled Release》;20120522;第161卷;第934页左栏第3段至第4段及右栏第2.2节;第936页左栏第3.1节;图1 *
微针在经皮给药及经皮疫苗转运中的应用;姜建芳 等;《医药导报》;20061231;第25卷(第10期);第1083页左栏倒数第一段及右栏第5段和第7段 *

Also Published As

Publication number Publication date
CN104706626A (en) 2015-06-17

Similar Documents

Publication Publication Date Title
CN104706626B (en) A kind of microneedle patch for being easy to animal vaccine to be administered and preparation method thereof
TWI554289B (en) Embeddable patch for transdermal drug delivery and method of manufacturing the same
CN111388407B (en) Microneedle array based on dopamine gel and preparation and application thereof
TWI528975B (en) Microneedle trandermal delivery device and microneedle transdermal delivery method using the same
CN104027324B (en) Soluble microneedle vaccine patch and preparation method thereof
CN105078880A (en) Macromolecular soluble microneedle used for cutaneous penetration of polypeptide and protein medicines and preparation method of macromolecular soluble microneedle
KR101747099B1 (en) Method of Preparing Micro-Needle Using Biocompatible Polymer
KR101931845B1 (en) Microneedle deposition technique
CN106474620A (en) A kind of polymer micro needle of medicine controlled release, preparation method and microneedle patch
CN112023033B (en) Two-section micro-needle array patch for simultaneously realizing BCG vaccine inoculation and diagnosis and preparation method thereof
CN101657188A (en) Solid drugs and vaccine dose
WO2011071287A2 (en) Ìicroneedle for improving the absorption rate of an active agent
CN106902453B (en) A kind of needle body is solvable and the insoluble microneedle preparation method of substrate
CN205360022U (en) A polymer soluble micropin that is used for polypeptide and protein drug transdermal to dose
CN106039552A (en) Bubble-type microneedle and preparation method therefor
CN114917465A (en) Self-heating microneedle drug-loaded patch and preparation method thereof
JP2013094662A (en) Microneedle welding method
CN108464967A (en) A kind of biological needle and preparation method thereof for subcutaneous medicament controlled release
KR101779393B1 (en) Microneedle array comprising nucleic acid and manufacture method thereof
CN110840823A (en) A kind of transfersome composite autolyzed microneedle and preparation method thereof
CN109106682A (en) A kind of painless percutaneous dosing microneedle patch of biological absorbable and preparation method thereof
CN108478520A (en) A kind of coating microneedle array and preparation method thereof accurately controlling drugloading rate
WO2018047800A1 (en) Double-needle type microneedle
CN114762675B (en) Controlled release type rabies vaccine soluble microneedle
CN110193082A (en) A kind of preparation method of sterile rabies vaccine coating micropin

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant